JP6713001B2 - Pharmaceutical composition containing silybin, VE, and L-carnitine - Google Patents

Description

The present invention relates to the field of medicine, particularly to a drug composition containing silybin for treating liver diseases.

In the late 60's and 80's of the 20th century, H.I. A West German pharmacist represented by wagner extracted and separated the active ingredient from the fruit of Silybummarianum, a plant of the genus Milk Thistle, and called it silymarin. This is a novel flavone compound having a C-9 substituent, that is, a flavonolignan formed by condensing flavanonol and a phenylpropanoid derivative. Silybin (Silibin) is one of the main components of Milk Thistle. Pharmacological and toxicological studies have revealed that it has significant effects in protecting and stabilizing the hepatocyte membrane, promoting hepatocyte recovery and improving liver function. Each has a different degree of protection and therapeutic action against liver damage caused by various toxins such as carbon tetrachloride, thioacetamide, muscarine, phalloidin, and retrocine. It is available for acute and chronic hepatitis, early cirrhosis, fatty liver, addictive or drug-induced liver disease.

Since silybin is poorly soluble in water and general organic solvents, it is poorly absorbed by oral administration, its bioavailability is low, and its clinical effect is affected. In order to improve its bioavailability, domestic and foreign pharmacy workers have conducted a lot of research. Countermeasures for improving absorption of poorly soluble drugs usually include ultrafine pulverization, chlorination, addition of solubilizing agents, and the like. In recent years, methods such as preparation of cyclodextrin inclusion compound, solid dispersion, synthesis of phospholipid complex, and preparation into different dosage forms have been adopted, and research shows that solubility and bioavailability are significantly improved. Has been done.

From the point of view of solid formulations, phospholipid complexes are special solid dispersions, have a fixed melting point, stable chemical properties, and are different from molecular compounds (complexes) of drugs and phospholipids. .. This type of compound depends on the type of phospholipid and the difference in the ratio of drug to phospholipid, and one phospholipid molecule may be bound to a different number of drug molecules. Due to the spectroscopic properties of the complex, the strong interaction between the drug and the polar group part of the phospholipid suppresses the free rotation of the single chain in the molecule, while the two long-chain fatty acid chains of the phospholipid are complexed. It is speculated that it does not participate in, is freely mobile and covers the polar portion of the phospholipids to form one lipophilic surface, making the complex highly lipophilic. This changes the physicochemical properties of the drug, increasing its lipophilicity and decreasing its water solubility. It promotes the binding between the drug molecule and the cell membrane, promotes absorption, and improves the bioavailability of the drug. Pu'er tea is a local tea unique to Yunnan. The climate of the production area is mild, the rainfall is sufficient, and the fog is covered. Pu'er tea consists of dried green tea of the large leaf species of the origin of Yunnan. There are two series. There are two types of shape, green tea and pressured tea, which have a unique quality in which the natural ripening process continues to proceed even after the finished product, and the more it matures, the deeper the taste.

Pu'er tea is the only post-fermentation type tea, and its harmful substances such as theophylline and polyphenols are differentiated in the fermentation process for a long period of time, so the quality is gentle and not irritating to the human body. It can also accelerate metabolism and accelerate the elimination and metabolism of fats and toxins in the body. For problems such as obesity, hypertension, hyperglycemia, and hyperlipidemia that are currently bothering people living in the city, Pu'er tea is, for example, detox, gastric recuperation, anti-inflammatory, cholesterol reduction. It can exert excellent relaxation effects such as eliminating cellulite and removing lipids, beauty and diet. According to modern technology, puer tea has effects such as improving insulin resistance, regulating blood lipid and leptin levels, and blocking the accumulation of fat in liver parenchymal cells due to insulin resistance in the body to some extent. It has been revealed that it has an action such as.

Non-alcoholic fatty liver disease (NAFLD) is a metabolic stress-related liver injury closely associated with insulin resistance and genetic predisposition, and its pathological changes are alcoholic fatty liver. Similar to a disease. The lesion body is located in the hepatic lobule and is characterized by fatty degeneration of hepatocytes and accumulation of fat, but it is a clinicopathologic syndrome with no excessive drinking history. NAFLD represents liver lesions of varying degrees, from simple fatty liver with no inflammation to severe inflammatory reactions of marked fibrosis and thus to cirrhosis, mainly in simple fatty liver, steatohepatitis, and fatty cirrhosis. Including two types.

Method for treating non-alcoholic fatty liver disease 1. Prevent and treat primary disease or related risk factors.
2. Basic treatment: Decide rational energy intake, adjust food and drink composition, perform moderate aerobic exercise, and correct poor lifestyle and behavior.
3. Avoiding Exacerbation of Liver Damage: Preventing rapid weight loss, drug abuse, and other factors that can lead to exacerbation of liver disease.
4. Diet: Overweight, visceral obesity, and NAFLD patients who rapidly gain weight in a short period of time all need to control their weight and reduce their waist by changing their lifestyle. Those who have monthly weight loss <0.45 kg or body weight index (BMI)> 27 kg/m ² and abnormalities of two or more indicators such as blood lipids, blood sugar, blood pressure, etc. in 6 months of basic treatment, It may be considered to take an additional diet drug such as sibutramine or orlistat, and the weekly weight loss should preferably not exceed 1.2 Kg (for children, it does not exceed 0.5 Kg each week), and BMI> For patients with obesity-related diseases such as 40 kg/m ² or BMI>35 kg/m ² and sleep apnea syndrome, consider gastric bypass surgery diet (II-1, II-2, II-3, III). May be.
5. Insulin sensitizer: Improvement of insulin resistance and control of blood glucose (II-1, II-2, II-3) in people with type 2 diabetes, impaired glucose tolerance, increased fasting blood glucose, and visceral fat obesity In order to achieve the above, the drug application of metformin and thiazolidinediones may be considered.
6. Hypolipidemic: Abnormal blood lipids continue to present mixed hyperlipidemia or hyperlipidemia, even after basic treatment for 3 to 6 months or longer and/or on diet antidiabetic drugs. People with more than one risk factor should be considered to take additional lipid-lowering drugs (II-1, II-2, II-3) such as fibrate, statin, or probucol ..
7. Liver disease remedy: Liver dysfunction with NAFLD, metabolic syndrome, ineffective after 3-6 months of basic treatment, and liver biopsy demonstrated NASH, and the course of the disease shows a chronic progressive course May aid in treatment with liver disease therapeutic agents to promote antioxidant, anti-inflammatory and anti-fibrosis. Based on drug performance and disease activity and stage, polyenephosphatidylcholine, vitamin E, silymarin. , And related drugs such as ursodeoxycholic acid (II-1, II-2, II-3, III) may be reasonably selected and taken, but multiple types of drugs should not be taken at the same time. Absent.
8. Liver transplantation: Mainly used to treat patients with end-stage liver disease related to NASH and some idiopathic cirrhosis and liver dysfunction, but metabolic status (III) should be examined before liver transplantation. BMI> 40 kg/m ² is a contraindication to liver transplantation (III).

As for the above-mentioned treatment method, there is not yet a method in which a plurality of types of treatment methods are mixed and used, such as a method in which a hypoglycemic agent and a liver disease drug are used in combination, or a lipid lowering and a liver disease agent are used in combination. Therefore, it is an urgent task to find a drug that can have multiple types of health improving functions.

In order to solve the above technical problems, the present invention provides a drug composition having a therapeutic effect on non-alcoholic fatty liver and a preparation thereof.

The present invention provides a method for preparing the drug composition and its formulation.

The present invention is realized by the following techniques.

A pharmaceutical composition, which is prepared from the following weight ratio of crude drug.
Silybin 8.75-60 parts Phospholipid 15-65 parts Pu'er tea extract 25-200 parts Vitamin E 6.25-40 parts L-carnitine 8.3-60 parts

The above is preferably prepared from the following weight ratio crude drug.
Silybin 25-40 parts Phospholipid 30-50 parts Pu'er tea extract 80-120 parts Vitamin E 10-30 parts L-carnitine 25-40 parts

Most preferably, the above is made from a crude drug in the following weight ratios.
Silybin 35 parts Phospholipids 42 parts Pu'er tea extract 100 parts Vitamin E 16.7 parts L-carnitine 33.3 parts

The phospholipid described in the present invention is soybean phospholipid or lecithin mainly containing phosphatidylcholine, and soybean phospholipid is preferable.

The action of phospholipids in the present invention is to promote dissolution and absorption of drugs. Silibin belongs to a poorly soluble and low membrane permeable drug, but phospholipids can improve silybin solubility after binding with silybin to form a phospholipid complex, thus improving drug bioavailability. Let

The above-mentioned silybin and phospholipid are either obtained by conventional techniques or commercially available. In order to exert the therapeutic effect of the present invention better, the silybin of the present invention is preferably prepared by the following method. That is, each silymarin is dissolved in 80% ethanol, filtered, the precipitate is dissolved 3 times using 95% ethanol, and the remaining precipitate is collected. The precipitate is dissolved in absolute ethanol, filtered, and a certain amount of water is added to the filtrate to precipitate the precipitate. The precipitate is collected by filtration, and the precipitate is collected under reduced pressure, dried, ground, and mixed. Good.

The Pu'er tea extract is commercially available, and Deepur's Pu'er powdered tea is preferred. Although it may be prepared according to the prior art, in order to exert the therapeutic effect of the present invention better, Pu'er powdered tea or an extract of Pu'er tea is prepared according to Patent Publication Nos. It is preferably prepared according to CN101961059B.

1 illustrates the process for producing Pu'er's powdered tea.
Step 1: Pour tea leaves in water and brew for 2-4 extractions. Extraction is performed for 0.5 to 2 hours each time, 6 to 12 times volume of water is added, the extract is filtered, and the filtrate is treated under the condition of ≦70° C., the weight of the tea leaves: the volume of the concentrate = 1 : Concentrate under reduced pressure until 2-1:3.
Step 2: Centrifuge the concentrate using a centrifuge, concentrate the centrifuge under reduced pressure at 45 to 65° C. until the specific gravity becomes 1.1 to 1.25, and perform spray drying or microwave drying. Done to get a concentrated cream.

The preferred steps are as follows.
Step 1: Pour tea leaves in water, boil vigorously, brew and extract 3 times. Extraction is performed for 0.5 to 2 hours each time, 6 to 12 times volume of water is added, the extract is filtered, and the filtrate is treated under the condition of ≦70° C., the weight of the tea leaves: the volume of the concentrate = 1 : Concentrate under reduced pressure until 2-1:3.
Step 2: Centrifuge the concentrated solution using a tripod centrifuge, further centrifuge the resulting separated solution using a tube centrifuge, and centrifuge solution at a specific gravity of 1. Concentrate under reduced pressure to 1 to 1.25 and perform spray drying or microwave drying to obtain a concentrated cream.
Here, the conditions of the tube type centrifugal separation are the number of rotations of the centrifuge: 15,000 to 19,000 rotations/min, and the spray drying conditions are a blast temperature: 140 to 190° C. and a wind discharge temperature: 75 to It is 95°C.

The most preferred steps are as follows.
Pour tea leaves in water, boil vigorously, brew and extract 3 times. The first time is decocted for 1.5 hours, 10 times volume of water is added, the second time is decocted for 1.5 hours, 8 times volume of water is added, the third time is decocted for 1 hour, and 8 times volume of water is added. The extract is filtered, and the filtrate is concentrated under reduced pressure at a temperature of ≦70° C. until the weight of tea leaves:volume of concentrate=1:2 to 1:3. The concentrate is centrifuged using a tripod centrifuge, the centrifuge is further centrifuged using a tube centrifuge, and the centrifuge at 45 to 65°C has a specific gravity of 1.1 to 1.25. Concentrated under reduced pressure until sprayed, and spray-dried or microwave-dried to obtain a concentrated cream.
Here, the conditions of the tube type centrifugal separation are the number of rotations of the centrifuge: 15,000 to 19,000 revolutions/min, and the spray drying conditions are the air supply temperature: 140 to 190° C. and the air discharge temperature: 75. ~95°C.

The vitamin E is any one of commercially available vitamin E, vitamin E acetate, and vitamin E succinate that can be used for nutritional support or medicinal purposes.

The L-carnitine is any one of commercially available L-carnitine and L-carnitine tartrate which can be used for nutritional support or medicinal purposes.

The above composition is blended on the basis of weight, and may be increased or decreased according to the corresponding ratio when producing, in the case of large-scale production, in kg, or t (ton) May be used as a unit, and g may be used as a unit for small-scale drug preparation. The weight may be increased or decreased, but the ratio of the weight ratios of the galenical materials between each composition does not change.

The above weight ratios have been obtained by scientific selection, and even for special sick people such as those who are severely or mildly ill, obese or thin, even if the corresponding composition ratios are adjusted. Well, if the increase or decrease does not exceed 10%, the effect of the drug is essentially unchanged.

When preparing a drug preparation, it can be prepared into any medicinal dosage form, and such dosage forms include tablets, coated tablets, film-coated tablets, enteric coated tablets, capsules, hard capsules and soft capsules. , Oral liquids, lozenges, granules, pills, powders, salves, potions, suspensions, solutions, injections, suppositories, ointments, plasters, creams, sprays, and patches To be elected. Oral dosage forms are preferred, with tablets, capsules and granules being most preferred.

The composition of the present invention may optionally contain an acceptable carrier for some drugs, and adopts the conventional technology of pharmacy such as mixing a drug active substance and an acceptable carrier for the drug. Then, the drug preparation may be prepared. The carrier acceptable for the drug includes mannitol, sorbitol, sorbic acid or potassium sorbate, sodium disulfite, sodium bisulfite, sodium thiosulfate, cysteine hydrochloride, thioglycolic acid, methionine, vitamin A, vitamin C, Vitamin E, vitamin D, Azone, disodium ethylenediaminetetraacetate, calcium disodium ethylenediaminetetraacetate, monovalent alkali metal carbonates, acetates, phosphates or their aqueous solutions, hydrochloric acid, acetic acid, sulfuric acid, phosphoric acid, Amino acids, sodium chloride, potassium chloride, sodium lactate, xylitol, maltose, glucose, fructose, dextran, glycine, starch, sucrose, lactose, mannitol, silicon derivatives, cellulose and its derivatives, alginates, gelatin, polyvinylpyrrolidone, glycerol, propylene. Glycol, ethanol, Tween 60-80, span-80, beeswax, lanolin, liquid paraffin, 1-hexadecanol, gallic acid esters, agar, triethanolamine, basic amino acids, urea, allantoin, calcium carbonate, bicarbonate It is selected from calcium, surfactant, polyethylene glycol, cyclodextrin, β-cyclodextrin, phospholipids, kaolin, talc, calcium stearate, magnesium stearate and the like. The carrier is one or more of microcrystalline cellulose, lactose, starch, sodium carboxymethyl cellulose, low-substituted hydroxypropyl cellulose, and talc.

When the composition of the present invention is prepared as a drug, a unit dose of the drug may contain 0.1 to 1000 mg of the drug active substance of the present invention, and the rest may be a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier may be 0.1 to 99.9% by weight of the total weight of the formulation. Preferably, the pharmaceutically acceptable carrier may be 40-70% by weight of the total weight of the formulation.

When using the Chinese herb composition or preparation of the present invention, the usage and dose can be determined according to the condition of the sick person.

The drug composition formulation according to the present invention is a slurry method, the number of revolutions is 100 rpm, the temperature is 37° C. under the elution conditions, and the dissolution medium is 1000 ml of hydrochloric acid solution of pH 1.2. , Granules, etc., and the dosage is 1 capsule/1 tablet/1 bag of granules. Regarding the dissolution rate outside the body, the cumulative dissolution rate for 2 hours is 60% or more, and the dissolution rate after 30 minutes is 15% or more.

The method for preparing the drug composition of the present invention includes the following steps.
a) preparing a prescribed amount of raw material,
b) Prescription amounts of silybin and phospholipids are weighed and dissolved in absolute ethanol, heated under reflux to clarify the solution, and after continuing heating for a certain period of time, the clear solution is concentrated under reduced pressure to a certain volume. Then, a step of preparing the silybin complex solution to obtain the silybin complex solution,
c) Weigh a prescribed amount of Pu'er tea extract as a base material, use the silybin complex solution prepared in b) as a liquid material, and prepare granules by a fluidized fluid injection method of a fluidized bed, and inject the entire liquid composition. Granulation step, drying after injection,
d) A total mixing step of uniformly mixing a prescribed amount of vitamin E and L-carnitine, and then uniformly mixing the mixture of both with the granules of step c) according to an increasing amount method to obtain a drug composition. ,
The present invention further comprises a pharmaceutical preparation step e), wherein the drug composition obtained in the above step d) and a pharmaceutically acceptable carrier are prepared into a pharmaceutically acceptable dosage form according to a usual pharmaceutical preparation process. ..

It is further preferred that the preparation method of the present invention includes the following steps.
a) preparing a prescribed amount of raw material,
b) Prescribed amounts of silybin and phospholipid are weighed and dissolved in absolute ethanol, heated and refluxed to clarify the solution, and after continuing heating for a certain period, until the clear solution has a certain volume. Concentrating under reduced pressure to obtain a silybin complex solution, a step of preparing the silybin complex solution,
c) Weigh a prescribed amount of Pu'er tea extract as a base material, use the silybin complex solution prepared in b) as a liquid material, and prepare granules by a fluidized fluid injection method of a fluidized bed, and inject the entire liquid composition. Granulation step, drying after injection,
d) A total mixing step of uniformly mixing a prescribed amount of vitamin E and L-carnitine, and then uniformly mixing the mixture of both with the granules of step c) according to an increasing amount method to obtain a drug composition. ,
e) A step of preparing a drug for preparing a usual preparation from the drug composition and a pharmaceutically acceptable carrier.
Here, the heating time described in step b) is 0.5 to 1.5 hours, the concentrated volume is 5% to 20% of the original volume, and the vacuum concentration temperature is 60 to 80°C.
Here, the parameters of the fluidized bed in step c) are such that the temperature of the material is 40 to 65° C., and parameters such as the number of revolutions of the fan, the air supply temperature, and the liquid injection frequency are adjusted in the granulation process, Keeps the material in a good fluidized state.
After the completion of granulation, it is dried for 10 to 60 minutes, and the drying temperature is 55 to 65°C.

The effects of Pu'er tea, such as improving insulin resistance and adjusting blood lipid and leptin levels, can block the accumulation of liver parenchymal fat due to insulin resistance in the body to some extent, and at the same time, it can also reduce the radical group of silybin. Synergistic with the strong ability of scavenging and preventing oxidative stress, both have good anti-NAFLD (non-alcoholic fatty liver disease) effect.

Increasing Vitamin E (Ve) further improves the beauty and facial effects of the product, and increasing L-carnitine enhances the lipid lowering and diet effects of the product.

Test Example 1 In vitro dissolution test

The dissolution rate of the silybin-phospholipid-puer tea-vitamin E-L-carnitine composition produced in Examples 16 to 20 was measured under the following conditions. The choice of elution method is based on the nature of silybin, the main component in the composition, which belongs to the fourth class in the classification system of biopharmaceuticals (BCS), which has poor solubility and low membrane permeability. It is a drug, and dissolution dissolution and absorption are both rate limiting steps. Therefore, improved bioavailability of a drug should be addressed simultaneously from both improved drug dissolution and improved absorption. The dissolution step of silybin is mainly performed in the stomach, the absorption step is mainly performed in the small intestine, and the detection of the dissolution rate of the drug in vitro contributes to the improvement of the bioavailability of the drug. .. Therefore, the following dissolution method is selected to evaluate the dissolution of the composition. In the slurry method, the rotation speed is 100 rpm, the temperature is 37° C., the dissolution medium is 1000 ml of hydrochloric acid solution having a pH of 1.2, and the dosage is 1 capsule/1 tablet/1 bag of granules. The sampling time points are 15, 30, 45, 60, 90 and 120 minutes, respectively. The cumulative solubility is measured. The results are shown in Table 1 below.

The dissolution was measured for the comparative preparation (silibin-phospholipid complex preparation, manufactured by the laboratory) and compared with the silybin-phospholipid-puar tea-vitamin E-L-carnitine composition prepared in Examples 19 to 23, The results are shown in Figure 1. The following contents can be understood from the data of Table 1 and the graph of FIG.

The silybin-phospholipid-puar tea-vitamin E-L-carnitine composition produced by employing the preparation method of the present invention is significantly superior to the silybin-phospholipid complex of the comparative preparation in terms of in vitro release. Surprisingly, the cumulative solubility of the composition in a hydrochloric acid solution having a pH of 1.2 for 2 hours reached 80% and was almost completely dissolved, which is a double increase as compared with the solubility of the comparative formulation. It solves the problem of low solubility and low bioavailability of silybin, and provides a basis for subsequent dosage setting of the silybin composition and study of its safety and efficacy in the body.

Summarizing the data of in vitro dissolution test and in vivo efficacy study, the present invention improves the absorption of the drug by complexing silybin and phospholipid to improve the affinity of the drug and the biological membrane, and By combining the silybin-phospholipid complex with puer tea, vitamin E, L-carnitine to improve the dissolution of the drug, the bioavailability of the main component silybin is improved both from dissolution and absorption improvement.

Test Example 2 Internal efficacy test

1 Experimental Animal 80 6-week-old male C57 BL/6J leptin-deficient mice (ob/ob) with SPF levels and 10 6-week-old male C57 BL/6J (ob/m) mice with SPF levels Is provided by Beijing Huafeng Biological Science and Technology Co., Ltd. and is bred in a barrier animal room of the Pharmacology and Toxicology Research Center of the Tenshi Research Institute, the temperature is 20°C to 25°C, and the relative humidity is 60%. Five mice were placed in each cage, the lighting time was 12 hours, and the feed was added in a fixed amount at a fixed time. The ob/ob mice were fed with a high-fat diet (HFD, D12492), and C57 BL/6J mice. Is fed with ordinary feed, and both are provided by Beijing Huafu Kang Biological Science and Technology Co., Ltd., and they are allowed to drink water freely and exchange bedding daily.

2 Test Product The silybin phospholipid complex is provided by Tasley Pharmaceutical Group Company Limited, lot number is 50090203, and Pu'er's powdered tea is a tan powder, Tasley Pharmaceutical Group Company. Provided by Limited, lot number is Z001 PE(2014)C06(H), L-carnitine tartrate (containing 68.4% L-carnitine) is provided by Tohoku Pharmaceutical Group Co., Ltd., lot number Is 0661504001, water-soluble vitamin E is produced by sigma company, and all the above test samples are stored in the sample shelf of the sample room provided by the Institute of Pharmacology and stored at room temperature while avoiding light. ..

3 Experimental method

3.1 Experimental Design and Packet The silybin phospholipid complex was prepared by calculating the dosage of the experimental animal based on the daily dose of 3 g of human (containing 420 mg of silybin and 504 mg of soybean phospholipid), and extracted from Pu'er tea. The dose of the experimental animal was calculated on the basis of 1.2 g of the daily dose of a human, the compounding ratio of 5 different compositions, and the experimental dose design are shown in Table 2. Therefore, the dose of each of the test animals was set to the dose of the clinically equivalent effect of the corresponding test substance, and the calculation formula is as follows.
Animal Experiment Dosage = Recommended Human Dosage/60kg*12.3

3.2 Administration of Test Product After rearing for 1 week adaptively, 80 6-week-old ob/ob mice were randomly divided into 8 groups, and model group, silybin phospholipid complex group, composition 1 group, composition There are 2 sets, 3 sets, 4 sets, and 5 sets, and each set has 10 animals. In addition, 10 6-week-old C57BL/6J mice are a normal group. The normal group of mice is fed with a normal diet, and both the model group and the dosing group are fed with a high fat diet (HFD, D12492). For mice of different drug interference groups, corresponding doses of drugs were intragastrically administered, and the doses of the five compositions are shown in Table 1. In each case, the same amount of distilled water is administered and the gastric administration is performed for 6 consecutive weeks.

Mice are allowed free access to food, water, and are weighed weekly and the dosage adjusted based on body weight within the experimental period. After the last administration, the animals were fasted only for 12 hours after watering, weighed, and the eyeball was removed to collect blood. Then, the mouse was killed by cervical dislocation, and the liver was rapidly dissected to remove the physiological saline. Wash with water, absorb water with filter paper, weigh and store in a refrigerator at -20°C.

3.3 Detection index and method

3.3.1 Observation of normal conditions During the experimental period, each group of mice is weighed once a week.

3.3.2 Calculating liver index and observing general morphology of liver After the experiment was completed, the liver was weighed and the liver index was calculated, liver index (%)=wet weight of liver/body weight*100% Is.

3.3.3 Detection of Serum Biochemical Index For all mice, the eyeball was removed, blood was collected, centrifuged at 3000 r/min for 15 minutes, serum was separated and collected in an EP tube. And store in a refrigerator at -20°C. The 7020 biochemical automated analyzer is used to detect the content of aspartate aminotransferase (AST), alanine aminotransferase (ALT), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C) in serum.

3.3.4 Insulin Resistance Index The Elisa kit is used to detect serum FINS and calculate the insulin resistance index by the formula.

3.3.5 Pathological Detection of Liver Tissue Frozen liver tissue is taken to prepare a frozen slice, and oil red O staining is used to observe the degree of degeneration of liver fat. The operation steps for oil red O staining are: frozen slices → thoroughly wash with distilled water → stain with oil red O diluted solution for 10 to 15 minutes → remove 6 ml of oil red O saturated solution and 4 ml of distilled water Add, add, leave for 5-10 minutes, filter, and then use → Differentiate with 60% ethanol until the substance between them becomes clear under the microscope → Wash with water → Stain the nucleus with hematoxylin/eosin → Wash with water → Seal the slice with neutral rubber → Observe with a microscope.

3.4 Data processing As for the statistical method, analysis was performed using SPSS 15.0 statistical software, the data were shown as mean ± standard difference, and t-test was adopted, before and after treatment of each group, and between groups. Each item was analyzed and compared to observe the presence or absence of a difference in the index, and P<0.05 was regarded as a difference, which was statistically significant.

4 experimental results

4.1 Effect of Each Test Product on Body Weight Each group of mice is weighed once a week during the experimental period, and the effect of each group of drugs on the body weight of non-alcoholic fatty liver is observed. As shown in Table 3, the weight gain of the normal group of mice is slow, and the weight gain of the model group of mice is fast. After 6 weeks of administration, each of the other administration groups, except the silybin phospholipid complex group, was able to suppress the weight gain of mice to a different degree (P<0.05), and the silybin phospholipid complex, puer tea extract , L-carnitine, VE co-utilization is clearly superior to using the silybin phospholipid complex alone.

4.2 Effect of each test substance on the liver index As shown in Table 4, the body weight, the liver wet weight, and the liver index of the mouse of the model group were clearly increased as compared with the normal group (P<0. .01), the silybin phospholipid complex can reduce liver wet weight in mice (P<0.05), but has no significant effect on mouse body weight and liver index (P>0.05). ), all of the five compositions can reduce liver wet weight and liver index in mice to different extents (P<0.05), and the effect of the four synergistic groups is the silybin phospholipid complex. Is clearly superior to using alone.

4.3 Effect of Each Test on Blood Lipid, Liver Function, and Insulin Resistance Index As shown in Table 5, serum TC, LDL, ALT, AST of non-alcoholic fatty liver model mice compared to the normal group. The insulin resistance index was clearly increased (P<0.05), and the silybin phospholipid complex had no apparent improvement for each index that was abnormally increased (P>0.05). Combinations of different ratios of silybin phospholipid complex, puer tea extract, VE, L-carnitine, except composition 4 had no apparent improvement in serum ALT (P>0.05), each other composition was All of them can significantly reduce TC, LDL-C, ALT, AST, and insulin resistance index (P<0.05), and the effect is superior to that of using the silybin phospholipid complex alone.

4.4 Effect of Each Test on Liver Pathology in Mice Oil Red O staining was mild or moderate depending on the size and number of red granules in hepatocytes under the light microscope of Oil Red O staining of frozen tissues of the liver. , Severe, divided into three. Mild, that is, 1/3 to 2/3 of red granules per unit area under an optical microscope were evaluated as 1 point, and moderate, that is, red granules in hepatocytes of 2/3 or more were found. If it is included, it is evaluated as 2 points, if it is severe, that is, in all almost all hepatocytes, there are red granules, it is evaluated as 3 points, and if no fatty degeneration is observed, it is evaluated as 0 point.

As shown in Table 6, in the liver tissues of the model group, almost all hepatocytes had steatosis, and the pathological evaluation score was clearly higher than that of the normal group (P<0.01. ), when using the silybin phospholipid complex or puer tea extract alone, no clear improvement was observed in the pathological evaluation score of the liver (P>0.05), and the silybin phospholipid complex and puer tea extract were obtained at different ratios. , VE, and L-carnitine can be used to remarkably improve hepatic steatosis and reduce the pathological evaluation score (P<0.05). The effect is silybin-phospholipid complex, puer Better than using tea extract alone.

5 Experimental conclusion As can be seen from the above experimental results, the body weight, liver index, blood lipid, ALT, AST, and insulin resistance index of the mice of the non-alcoholic fatty liver model group were all significantly higher than those of the blank group. , Liver tissue is severely fatty modified. Pu'er tea can improve insulin resistance and regulate blood lipid, and L-carnitine also has a good blood lipid regulating effect, and at the same time, has a strong ability to remove radical groups and prevent oxidative stress. There is a clear improvement in hepatic steatosis when used in combination with the four in combination with potent silybin and VE.

Figure 6 is an in vitro release graph, where each sample is the silymarin comparative formulation, silymarin without puer tea, silybin-phospholipid-puer tea composition prepared in Examples 16-20.

The present invention is further illustrated by the following specific examples, which do not limit the present invention.

Example 1
26.25 g of silybin, 45 g of soybean phospholipid, 75 g of puer tea extract, 18.75 g of vitamin E, and 25 g of L-carnitine are prepared.
a) Preparation of silybin complex solution: Prescription amounts of silybin and soybean phospholipid are weighed and dissolved in absolute ethanol, heated and refluxed to clarify the solution, and then heated for 1 hour, and recovered ethanol is recovered to the original volume. Concentrate under reduced pressure to 15% of.
b) Granulation: A prescribed amount of Pu'er tea extract is weighed and used as a base material, the silybin complex liquid prepared in a) is used as a liquid material, and granulated by a fluidized liquid injection method of a fluidized bed, and the temperature of the material is Is controlled at 40° C., and after the liquid composite is completely injected, it is dried at a drying temperature of 60° C. for 20 minutes.
c) First, the prescribed amounts of vitamin E and L-carnitine are uniformly mixed, and then the mixture of both and the granules prepared in step b) are uniformly mixed according to the method of increasing the volume by equal amounts, put in a bag, and then 1000 Make bag granules.

Example 2
180 g of silybin, 195 g of soybean phospholipid, 600 g of puer tea extract, 120 g of vitamin E, and 180 g of L-carnitine are prepared.
a) Preparation of silybin complex: Prescription amounts of silybin and soybean phospholipid are weighed and dissolved in absolute ethanol, heated and refluxed to clarify the solution, and then heated for 1.5 hours, and recovered ethanol is recovered. Concentrate under reduced pressure to 20% of the volume.
b) Granulation: A prescribed amount of puerh tea extract is weighed and used as a base material, the silybin composite liquid prepared in a) is used as a liquid material, and granulated by a fluidized fluid injection method of a fluidized bed, and the temperature of the material is The temperature is controlled to 65° C., and after the liquid composite is completely injected, it is dried at a drying temperature of 65° C. for 60 minutes.
c) First, the prescribed amounts of vitamin E and L-carnitine are uniformly mixed, and then the mixture of both and the granules prepared in step b) are uniformly mixed according to the method of increasing the volume by equal amounts, put in a bag, and then 1000 Make bag granules.

Example 3
26.25 g of silybin, 195 g of phospholipid, 600 g of puer tea extract, 18.75 g of vitamin E, and 25 g of L-carnitine are prepared.
a) Preparation of silybin complex: Prescription amounts of silybin and soybean phospholipid are weighed and dissolved in absolute ethanol, heated and refluxed to clarify the solution, and then continuously heated for 0.5 hours to recover recovered ethanol. Concentrate under reduced pressure to a volume of 5%.
b) Granulation: A prescribed amount of puerh tea extract is weighed and used as a base material, the silybin composite liquid prepared in a) is used as a liquid material, and granulated by a fluidized fluid injection method of a fluidized bed, and the temperature of the material is It is controlled at 50° C., and after the liquid composite is completely injected, it is dried at a drying temperature of 55° C. for 10 minutes.
c) First, the prescribed amounts of vitamin E and L-carnitine are uniformly mixed, and then the mixture of both and the granules prepared in step b) are uniformly mixed according to the method of increasing the volume by equal amounts, put in a bag, and then 1000 Make bag granules.

Example 4
26.25 g of silybin, 195 g of phospholipid, 75 g of puercha tea extract, 120 g of vitamin E, 180 g of L-carnitine are prepared and prepared according to the method of Example 1 to prepare 1000 bags of granules.

Example 5
180 g of silybin, 45 g of phospholipid, 75 g of puer tea extract, 18.75 g of vitamin E and 180 g of L-carnitine are prepared and prepared according to the method of Example 1 to prepare 1000 bags of granules.

Example 6
180 g of silybin, 45 g of phospholipid, 600 g of puer tea extract, 120 g of vitamin E, and 25 g of L-carnitine are prepared and prepared according to the method of Example 1 to prepare 1000 bags of granules.

Example 7
180 g of silybin, 195 g of phospholipid, 75 g of puer tea extract, 18.75 g of vitamin E and 180 g of L-carnitine are prepared and prepared according to the method of Example 1 to prepare 1000 bags of granules.

Example 8
Prepare 26.25 g of silybin, 48.75 g of phospholipids, 75 g of puer tea extract, 37.5 g of vitamin E, and 50 g of L-carnitine and prepare according to the method of Example 1 to prepare 1000 bags of granules.

Example 9
Prepare 26.25 g of silybin, 48.75 g of phospholipids, 300 g of puer tea extract, 37.5 g of vitamin E and 50 g of L-carnitine and prepare according to the method of Example 1 to make 1000 bags of granules.

Example 10
Prepare 52.5 g of silybin, 97.5 g of phospholipid, 300 g of puer tea extract, 9.375 g of vitamin E, and 12.5 g of L-carnitine, and prepare according to the method of Example 1 to make 1000 bags of granules. ..

Example 11
75 g of silybin, 90 g of phospholipid, 240 g of puercha tea extract, 30 g of vitamin E, and 75 g of L-carnitine are prepared and prepared according to the method of Example 1 to prepare 1000 bags of granules.

Example 12
90 g of silybin, 108 g of phospholipid, 270 g of puercha tea extract, 60 g of vitamin E, 90 g of L-carnitine are prepared and prepared according to the method of Example 1 to prepare 1000 bags of granules.

Example 13
Prepare 105 g of silybin, 126 g of phospholipid, 300 g of puer tea extract, 75 g of vitamin E, 100 g of L-carnitine and prepare according to the method of Example 1 to prepare 1000 bags of granules.

Example 14
Prepare 105 g of silybin, 195 g of phospholipid, 300 g of puer tea extract, 75 g of vitamin E, 100 g of L-carnitine and prepare according to the method of Example 1 to prepare 1000 bags of granules.

Example 15
120 g of silybin, 150 g of phospholipids, 360 g of puer tea extract, 90 g of vitamin E, 120 g of L-carnitine are prepared and prepared according to the method of Example 1 to prepare 1000 bags of granules.

Example 16
Prepare 105 g of silybin, 126 g of phospholipid, 300 g of puercha tea extract, 75 g of vitamin E, 226.6 g of L-carnitine tartrate and prepare according to the method of Example 1 to make 1000 bags of granules.

Example 17
Prepare 105 g of silybin, 126 g of phospholipid, 300 g of puercha tea extract, 82.3 g of vitamin E acetate ester, 100 g of L-carnitine and prepare according to the method of Example 1 to make 1000 bags of granules.

Example 18
Prepare 105 g of silybin, 126 g of phospholipid, 300 g of puer tea extract, 92.4 g of vitamin E succinate, and 226.6 g of L-carnitine tartrate, prepare according to the method of Example 1, and prepare 1000 bags of granules. Create.

Example 19
The granules of Example 8 are prepared, 500 g of microcrystalline cellulose and 64 g of sodium starch glycolate are added, uniformly mixed, and put into a capsule to prepare 1000 particles.

Example 20
The granules of Example 9 are prepared, 274 g of microcrystalline cellulose and 64 g of sodium starch glycolate are added, uniformly mixed, and put into a capsule to prepare 1000 particles.

Example 21
The granules of Example 10 are prepared, 297 g of microcrystalline cellulose and 32 g of sodium starch glycolate are added, uniformly mixed, and put into a capsule to prepare 1000 particles.

Example 22
The granules of Example 13 are prepared, 30 g of microcrystalline cellulose and 64 g of sodium starch glycolate are put therein, uniformly mixed and put into a capsule to prepare 1000 grains.

Example 23
The granules of Example 14 are prepared, 25 g of microcrystalline cellulose is put therein, they are mixed uniformly, and they are put into capsules to prepare 1000 grains.

Example 24
The composition of Example 8 is prepared, 400 g of lactose, 100 g of starch, and 64 g of sodium starch glycolate are added, uniformly mixed, and put into a capsule to prepare 1000 grains.

Example 25
The composition of Example 13 is prepared, 20 g of lactose, 10 g of talc, and 64 g of low-substituted hydroxypropylcellulose are added, uniformly mixed, and put into a capsule to prepare 1000 grains.

Example 26
The composition of Example 13 is prepared, 30 g of microcrystalline cellulose and 64 g of sodium starch glycolate are added, uniformly mixed and pressed into tablets to produce 1000 tablets.

Claims (11)

A drug composition for treating non-alcoholic fatty liver, diet or lowering lipids, which is prepared from the components in the following weight ratios.
Silybin 8.75-60 parts Phospholipid 15-65 parts Pu'er tea extract 25-200 parts Vitamin E 6.25-40 parts L-carnitine 8.3-60 parts The drug composition according to claim 1, which is prepared from the components in the following weight ratios.
Silybin 25-40 parts Phospholipid 30-50 parts Pu'er tea extract 80-120 parts Vitamin E 10-30 parts L-carnitine 25-40 parts The drug composition according to claim 1, which is prepared from the components in the following weight ratios.
Silybin 35 parts Phospholipids 42 parts Pu'er tea extract 100 parts Vitamin E 16.7 parts L-carnitine 33.3 parts The drug composition according to any one of claims 1 to 3, wherein the vitamin E can be replaced with vitamin E acetate or vitamin E succinate. The L- carnitine, the pharmaceutical composition according to any one of claims 1 to 3, wherein the Ru L- carnitine tartrate der. A pharmaceutical composition according to any one of claims 1 to 3, and a pharmaceutically acceptable carrier,
Formulation of drugs medicine biological acceptable carrier, characterized in that the Ru 0.1 to 99.9% der of the total weight of the formulation by weight. The carrier is mannitol, sorbitol, sorbic acid or potassium sorbate, sodium disulfite, sodium bisulfite, sodium thiosulfate, cysteine hydrochloride, thioglycolic acid, methionine, vitamin A, vitamin C, vitamin E, vitamin D. Azone, disodium ethylenediaminetetraacetate, calcium disodium ethylenediaminetetraacetate, monovalent alkali metal carbonates, acetates, phosphates or their aqueous solutions, hydrochloric acid, acetic acid, sulfuric acid, phosphoric acid, amino acids, sodium chloride, Potassium chloride, sodium lactate, xylitol, maltose, glucose, fructose, dextran, glycine, starch, sucrose, lactose, mannitol, silicon derivatives, cellulose and its derivatives, alginate, gelatin, polyvinylpyrrolidone, glycerol, propylene glycol, ethanol, tween. 60-80, span-80, beeswax, lanolin, liquid paraffin, 1-hexadecanol, gallic acid esters, agar, triethanolamine, basic amino acids, urea, allantoin, calcium carbonate, calcium bicarbonate, surfactant , claim 6 polyethylene glycol, cyclodextrin, beta-cyclodextrin, phospholipids, kaolin, talc, calcium stearate, magnesium stearate, characterized in including that one or one or more of the inside of the microcrystalline cellulose The preparation according to. The above-mentioned preparations include tablets, coated tablets, film-coated tablets, enteric coated tablets, capsules, hard capsules, soft capsules, oral solutions, lozenges, granules, pills, powders, salves, potions, suspensions. , solutions, injections, suppositories, ointments, plasters, creams, sprays, formulation according to claim 7, wherein any one der Rukoto in patch. A method for preparing the formulation according to claim 6, comprising the steps of: a) providing a prescribed amount of raw material;
b) Prescription amounts of silybin and phospholipids are weighed and dissolved in absolute ethanol, heated and refluxed to clarify the solution, and after continuing heating for a certain period of time, the clear solution is concentrated under reduced pressure until a certain volume is reached. Then, a step of preparing the silybin complex solution to obtain the silybin complex solution,
c) Weigh a prescribed amount of Pu'er tea extract as a base material, use the silybin complex solution prepared in b) as a liquid material, and prepare granules by a fluidized fluid injection method of a fluidized bed, and inject the entire liquid composition. Granulation step, drying after injection,
d) a total mixing step of uniformly mixing vitamin E, L-carnitine and the granules of step c) to obtain a drug composition, and e) a conventional formulation from the drug composition and a pharmaceutically acceptable carrier. A preparation method comprising the step of preparing a drug for preparing. The heating time in step b) is 0.5-1.5 hours, the concentrated volume is 5%-20% of the original volume, the vacuum concentration temperature is 60-80° C., and in step c) The parameters of the fluidized bed are such that the material temperature is 40 to 65° C., and parameters such as the number of revolutions of the fan, the air supply temperature, and the frequency of liquid injection are adjusted in the granulation process to ensure a good fluidization state of the material. 10. The method according to claim 9, wherein after the granulation is completed, the granulation is completed, the granulation is completed for 10 to 60 minutes, and the drying temperature is 55 to 65°C. The formulation according to the drug composition or claim 6 according to any one of claims 1 to 3, the treatment and / or diet of nonalcoholic fatty liver, in the preparation of a medicament for the lower descending lipids.